1. Field of the Invention
The present invention is to provide a method of double allele specific PCR, and more particularly, to provide a method of double allele specific PCR for SNP microarray.
2. The Prior Arts
There are many different types of genetic variation in the human genome; the most studied class of variant is the single nucleotide polymorphism (SNP). SNPs lying in protein-coding regions of the gene can alter the code for an amino acid sequence and may affect the function of a protein and the regulation of a gene expression so as to cause disease.
Recently, high-throughput microarray chip technology has been wildly used to detect individual differences, to perform genetic correlation analysis, to analyze linkage disequilibrium or to screen drug metabolism genes, etc. However, the high-throughput microarray chip technology contains some defects: for example, the pre-treatment step prior to array hybridization is quite time-consuming, tens of thousands of SNPs data obtained after array screening are difficult to statistically analyze, and the instruments and suppliers are very expensive. Another technology, real-time polymerase chain reaction, is also wildly used due to its high specificity, sensitivity and accuracy, but it requires high cost to design a probe containing specific modification for each SNP, which results in higher overall costs for detection, and this technology is not suitable for large-scale SNP genotyping detection. Therefore, an effective, rapid, large-scale detection technology for singe nucleotide variation is needed.